After inducing calcium deficiency, both the sham (S) and ovariectomized (O) groups were given a standard diet with calcium citrate tetrahydrate. The ovariectomized (O) group was further divided into four subgroups, each comprising eight rats, and these subgroups received a standard diet enriched with calcium citrate tetrahydrate. Table 1 displays the diet formulas. Throughout the 6-week intervention period, the rats had unrestricted access to the diets and deionized water.
The selection of daidzein and tempeh doses was informed by prior studies that established their beneficial impacts on the skeletal well-being of rodents and humans. Through a comprehensive analysis of aglycon content, specifically daidzein, glycitein, and genistein, it has been demonstrated that the optimal daily consumption of tempeh should be set at 250 g [35 (link)]. The dosage of daidzein at 10 mg/kg diet was chosen based on our previous research [27 (link),28 (link),36 (link),37 (link),38 (link)], and the quantity of tempeh corresponds to the daidzein content. In this study, the amount of tempeh was adjusted to the appropriate amount of pure daidzein. To incorporate 250 g/kg of tempeh flour and 10 mg/kg of daidzein into the AIN93M diets, we carefully adjusted the diets by replacing starch with 250 g and 10 milligrams of the corresponding ingredients. This approach ensured that the diets had the correct amount of tempeh flour or daidzein while being nutritionally consistent.
We determined the dosages of isoflavones by referencing prior studies that have documented their positive impacts on bone health in both rodents and humans [35 (link)]. Furthermore, long-term therapy with alendronate bisphosphonate at a dosage of 3 mg/kg/day has been shown to benefit fracture healing and bone remodeling in ovariectomized rats [39 (link)]. The dose of alendronate bisphosphonate was adjusted weekly after measuring the rats’ body weight.
The selection of daidzein and tempeh doses was informed by prior studies that established their beneficial impacts on the skeletal well-being of rodents and humans. Through a comprehensive analysis of aglycon content, specifically daidzein, glycitein, and genistein, it has been demonstrated that the optimal daily consumption of tempeh should be set at 250 g [35 (link)]. The dosage of daidzein at 10 mg/kg diet was chosen based on our previous research [27 (link),28 (link),36 (link),37 (link),38 (link)], and the quantity of tempeh corresponds to the daidzein content. In this study, the amount of tempeh was adjusted to the appropriate amount of pure daidzein. To incorporate 250 g/kg of tempeh flour and 10 mg/kg of daidzein into the AIN93M diets, we carefully adjusted the diets by replacing starch with 250 g and 10 milligrams of the corresponding ingredients. This approach ensured that the diets had the correct amount of tempeh flour or daidzein while being nutritionally consistent.
We determined the dosages of isoflavones by referencing prior studies that have documented their positive impacts on bone health in both rodents and humans [35 (link)]. Furthermore, long-term therapy with alendronate bisphosphonate at a dosage of 3 mg/kg/day has been shown to benefit fracture healing and bone remodeling in ovariectomized rats [39 (link)]. The dose of alendronate bisphosphonate was adjusted weekly after measuring the rats’ body weight.
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